目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快...目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快速扩增法（RACE）克隆其全长cDNA，应用生物信息学对其功能进行初步分析，应用一步法半定量RT—PCR检测其在AML患者及正常人中的表达。结果获得了家族性AML相关新基因FAMLF的cDNA全长，该基因定位于染色体1q31．3，cDNA全长2313bp，开放读码框（ORF）为249bp，编码82个氨基酸的蛋白质，含有信号肽，富含亮氨酸重复单位（LRR—SD22），内在固有无序结构域等功能区。Blast检索为功能未知的新基因，已被GenBank收录，并命名为FAMLF，核酸注册号为EF413001，蛋白质注册号为ABN58747。新基因FAMLF在AML患者中的表达明显高于正常人，差异有统计学意义（2．61±0．66 vs 0．97±0．51，P〈0．01）。结论成功获得一个家族性AML相关新基因FAMLF cDNA全长，FAMLF基因在AML中高表达，可能是具有一定生物功能的新基因。展开更多
Aquilaria si,iensis, a kind of typically wounding-induced medicinal plant with a great economical value, is widely used in the production of traditional Chinese medicine, perfume and incense. Coronatine-insensitive pr...Aquilaria si,iensis, a kind of typically wounding-induced medicinal plant with a great economical value, is widely used in the production of traditional Chinese medicine, perfume and incense. Coronatine-insensitive protein 1 (COI1) acts as a receptor in jasmonate (JA) signaling pathway, and regulates the expression of JA-responsive genes in plant defense. However, little is known about the COI1 gene in A, sinensis, Here, based on the transcriptome data a full-length cDNA sequence of COI1 (termed as AsCOM was firstly cloned by RT PCR and rapid amplification of cDNA ends (RACE) strategies. AsCOI1 is 2330 bp in length (GenBank accession No. KM 189194), and contains a complete open frame (ORF) of 1839 bp. The deduced protein was composed of 612 amino acids, with a predicted molecular weight of 68.93 kDa and an iisoelectric point of 6.56, and was predicted to possess b-box and ERRs domains. Combining bioinformatics prediction with subcellular localization experiment analysis, AsCOI1 was appeared to locate in nucleus. AsCOI1 gene was highly expressed in roots and stems, the major organs of agarvimod formation. Methyl jasmonate (MeJA), mechanical wounding and heat stress could significantly induce the expression level of AsCOI1 gene. AsCOI1 is an early wound -responsive gene, and it likely plays sonic role in agarwood formation. (c) 2015 Chinese Pharmaceutical Association and Institute of Materia Medica Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.展开更多
Bombyx mori cytoplasmic polyhedrosis virus （BmCPV） is one of the major viral pathogens for the silkworm. To date, the molecular mechanism of BmCPV invasion has been unclear. We cloned the full length complementary ...Bombyx mori cytoplasmic polyhedrosis virus （BmCPV） is one of the major viral pathogens for the silkworm. To date, the molecular mechanism of BmCPV invasion has been unclear. We cloned the full length complementary （c）DNA which encodes the ubiquitin-activating enzyme El-domain containing proteinl （UbE1DC1） ofBombyx mori by using suppression subtraefive hybridization （SSH） and rapid amplification of com- plementary （c）DNA ends （RACE）. The full-length eDNA of UbE1DClgene is 1 919 bp, consisting of a 100 bp 5＇ untranslated region, a 637 bp 3＇ untranslated region and an 1 182 bp open reading frame （ORF）, encoding a 393 amino acid protein. The protein contained the THiF_MoeB_hesA_family domain, an adenosine triphosphate binding site, which belongs to the family of ubiquitin-activating enzyme El. Reverse transcription - polymerase chain reaction analysis from the silkworm tissues, namely silk gland, hemo- cyte, fat body, gonad and midgut revealed that UbE1DC1 was expressed in all the five tissues. The real-time quantitative polymerase chain reaction analysis indicated that the relative expression of UbE1DC1 in the normal midgut was approximately 9.78-fold of that in the BmCPV-infected midgut. It is implicated that UbEIDCI may play an important role in the interaction between the host and BmCPV invasion.展开更多
文摘目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快速扩增法（RACE）克隆其全长cDNA，应用生物信息学对其功能进行初步分析，应用一步法半定量RT—PCR检测其在AML患者及正常人中的表达。结果获得了家族性AML相关新基因FAMLF的cDNA全长，该基因定位于染色体1q31．3，cDNA全长2313bp，开放读码框（ORF）为249bp，编码82个氨基酸的蛋白质，含有信号肽，富含亮氨酸重复单位（LRR—SD22），内在固有无序结构域等功能区。Blast检索为功能未知的新基因，已被GenBank收录，并命名为FAMLF，核酸注册号为EF413001，蛋白质注册号为ABN58747。新基因FAMLF在AML患者中的表达明显高于正常人，差异有统计学意义（2．61±0．66 vs 0．97±0．51，P〈0．01）。结论成功获得一个家族性AML相关新基因FAMLF cDNA全长，FAMLF基因在AML中高表达，可能是具有一定生物功能的新基因。
基金supported by grants from the National Natural Science Foundation of China (Nos. 31100220, 81173481 and 31000136)the Program for Xiehe Scholars in Chinese Academy of Medical Sciences & Peking Union Medical College (No. 282)the Innovative Team and Innovative Talents Project of the Ministry of Science and Technology of China
文摘Aquilaria si,iensis, a kind of typically wounding-induced medicinal plant with a great economical value, is widely used in the production of traditional Chinese medicine, perfume and incense. Coronatine-insensitive protein 1 (COI1) acts as a receptor in jasmonate (JA) signaling pathway, and regulates the expression of JA-responsive genes in plant defense. However, little is known about the COI1 gene in A, sinensis, Here, based on the transcriptome data a full-length cDNA sequence of COI1 (termed as AsCOM was firstly cloned by RT PCR and rapid amplification of cDNA ends (RACE) strategies. AsCOI1 is 2330 bp in length (GenBank accession No. KM 189194), and contains a complete open frame (ORF) of 1839 bp. The deduced protein was composed of 612 amino acids, with a predicted molecular weight of 68.93 kDa and an iisoelectric point of 6.56, and was predicted to possess b-box and ERRs domains. Combining bioinformatics prediction with subcellular localization experiment analysis, AsCOI1 was appeared to locate in nucleus. AsCOI1 gene was highly expressed in roots and stems, the major organs of agarvimod formation. Methyl jasmonate (MeJA), mechanical wounding and heat stress could significantly induce the expression level of AsCOI1 gene. AsCOI1 is an early wound -responsive gene, and it likely plays sonic role in agarwood formation. (c) 2015 Chinese Pharmaceutical Association and Institute of Materia Medica Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.
文摘Bombyx mori cytoplasmic polyhedrosis virus （BmCPV） is one of the major viral pathogens for the silkworm. To date, the molecular mechanism of BmCPV invasion has been unclear. We cloned the full length complementary （c）DNA which encodes the ubiquitin-activating enzyme El-domain containing proteinl （UbE1DC1） ofBombyx mori by using suppression subtraefive hybridization （SSH） and rapid amplification of com- plementary （c）DNA ends （RACE）. The full-length eDNA of UbE1DClgene is 1 919 bp, consisting of a 100 bp 5＇ untranslated region, a 637 bp 3＇ untranslated region and an 1 182 bp open reading frame （ORF）, encoding a 393 amino acid protein. The protein contained the THiF_MoeB_hesA_family domain, an adenosine triphosphate binding site, which belongs to the family of ubiquitin-activating enzyme El. Reverse transcription - polymerase chain reaction analysis from the silkworm tissues, namely silk gland, hemo- cyte, fat body, gonad and midgut revealed that UbE1DC1 was expressed in all the five tissues. The real-time quantitative polymerase chain reaction analysis indicated that the relative expression of UbE1DC1 in the normal midgut was approximately 9.78-fold of that in the BmCPV-infected midgut. It is implicated that UbEIDCI may play an important role in the interaction between the host and BmCPV invasion.
